Thermoresponsive cell culture supports

Patent metadata

Field	Value
Publication number	US-10550366-B2
Application number	US-201715499964-A
Country	US
Kind code	B2
Filing date	Apr 28, 2017
Priority date	Apr 8, 2011
Publication date	Feb 4, 2020
Grant date	Feb 4, 2020

Abstract

The present invention relates to a cell culture support including a substrate and a thermoresponsive polymeric blend layer, wherein the polymeric blend layer includes at least one thermoresponsive polymer and at least one network forming adhesion promoter. The present invention further relates a method of making a cell culture complex including: providing a substrate; blending at least one thermoresponsive polymer and at least one network forming adhesion promoter to provide a polymeric blend; applying a thin film of said polymeric blend to the substrate to provide a polymeric blend layer on the substrate; curing the polymeric blend layer on the substrate to provide a cell culture support; and depositing cells onto said cell culture support, wherein the cells may optionally further include medium, to provide a cell culture complex.

First claim

What is claimed is:

1. A method of making a cell culture complex comprising:
providing a substrate;
blending at least one thermoresponsive polymer and at least one network forming adhesion promoter to provide a polymeric blend, wherein the network forming adhesion promoter is a non-protein adhesion promoter and has functional amino or carboxylic acid groups;
applying a thin film of the polymeric blend to the substrate to provide a polymeric blend layer on the substrate;
curing the polymeric blend layer on the substrate to provide a cell culture support; wherein the cell culture support includes the at least one network forming adhesion promoter in the form of a cross-linked network that is bound to the substrate and that interlocks chains of the at least one thermoresponsive polymer;
depositing cells onto the cell culture support to provide a cell culture complex, and
allowing the deposited cells of the cell culture complex to culture to thereby form a cultured cell layer.

2. The method of claim 1 , wherein the cultured cell layer comprises cells further characterized as anchor dependent cells.

3. The method of claim 1 , wherein the cultured cell layer comprises cells further characterized as adhesive cells.

4. The method of claim 1 , wherein the cultured cell layer comprises cells selected from the group consisting of fibroblasts, myoblasts, myotube cells, corneal cells, vascular endothelial cells, smooth muscle cells, cardiomyocytes, dermal cells, epidermal cells, mucosal epithelial cells, mesenchymal stem cells, embryonic stem (ES) cells, induced pluripotent stem (iPS) cells, osteoblasts, osteocytes, chondrocytes, fat cells, neurons, hair root cells, dental pulp stem cells, β-cells, hepatocytes, and combinations thereof.

5. The method of claim 1 , wherein the step of depositing cells includes depositing the cells within a medium.

6. The method of claim 1 , wherein the step of applying the thin film of the polymeric blend is a step of spin-coating the thin film of the polymeric blend onto the substrate.

7. The method of claim 1 , wherein the polymeric blend layer is devoid of adhesive proteins.

8. The method of claim 1 , wherein the polymeric blend layer is devoid of plasma or e-beam treatment.

9. The method of claim 1 , further comprising a step of detaching the cultured cell layer.

10. The method of claim 9 , wherein the step of detaching the cultured cell layer is devoid of proteolytic enzymes or mechanical agitations.

Assignees

• Newby Bi Min
• Patel Nikul
• Cavicchia John
• Zhang Ge
• Univ Akron

Inventors

• Newby Bi-Min
• Patel Nikul
• Cavicchia John
• Zhang Ge

Classifications

• C12N2539/10

  Coating allowing for selective detachment of cells, e.g. thermoreactive coating · CPC title

• C12N5/0668

  Mesenchymal stem cells from other natural sources · CPC title

• C12N5/0665

  Blood-borne mesenchymal stem cells, e.g. from umbilical cord blood · CPC title

• C12N5/0666

  Mesenchymal stem cells from hair follicles · CPC title

• C12N2533/12

  Glass · CPC title